Apoptosis of cultured granulosa-lutein cells is reduced by insulin-like growth factor I and may correlate with embryo fragmentation and pregnancy rate

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Apoptosis of cultured granulosa-lutein cells is reduced by insulin-like growth factor I and may correlate with embryo fragmentation and pregnancy rate Eija Bencomo, M.Sc., Rosa Pérez, M.Sc., Mari-Francis Arteaga, Ph.D., Elisa Acosta, M.Sc., Oscar Peña, M.Sc., Lourdes Lopez, M.D., Julio Avila, Ph.D., Angela Palumbo, M.D., Ph.D. Fertility and Sterility Volume 85, Issue 2, Pages 474-480 (February 2006) DOI: 10.1016/j.fertnstert.2005.08.014 Copyright © 2006 American Society for Reproductive Medicine Terms and Conditions

FIGURE 1 Fluorescent micrograph. (A) Viable (red nucleus) and apoptotic (red nucleus and green cytoplasm) mural GL cells after 48 hours of culture without serum. (B) Cumulus cells after 48 hours of culture without serum. Granulosa-lutein cells after 48 hours of culture in serum-free medium of one patient who conceived (C) and of one who did not conceive (D). Cells are marked with propidium iodide (0.01 mg/mL) and with a fluorescent marker for activated caspases, caspACE-FITC-VAD-FMK (10 μM). Bar = 50 μm. Bencomo. Apoptosis in human granulosa-lutein cells. Fertil Steril 2006. Fertility and Sterility 2006 85, 474-480DOI: (10.1016/j.fertnstert.2005.08.014) Copyright © 2006 American Society for Reproductive Medicine Terms and Conditions

FIGURE 2 (A) The percentage of apoptosis after 48 hours of culture without serum of mural GL cells that were obtained from patients <38 years old and ≥38 years old (P=.015, Mann-Whitney U). (B) Dose–response of GL cell apoptosis to IGF-I (0.3, 10, and 30 nM; P=.043, Kruskal-Wallis). a,bMeans are statistically different (Mann-Whitney U with Bonferroni correction: P<.05). (C) Percentage of apoptosis of GL cells from patients who became pregnant in that IVF cycle and from patients who did not, after 48 hours of culture without serum (P=.006, Student’s t-test). Bencomo. Apoptosis in human granulosa-lutein cells. Fertil Steril 2006. Fertility and Sterility 2006 85, 474-480DOI: (10.1016/j.fertnstert.2005.08.014) Copyright © 2006 American Society for Reproductive Medicine Terms and Conditions

FIGURE 3 Linear correlation between the percentage of apoptosis after 48 hours of culture without serum of mural GL cells vs. percentage of embryo fragmentation (Pearson’s correlation test: n = 88, r = 0.214, P<.05). Bencomo. Apoptosis in human granulosa-lutein cells. Fertil Steril 2006. Fertility and Sterility 2006 85, 474-480DOI: (10.1016/j.fertnstert.2005.08.014) Copyright © 2006 American Society for Reproductive Medicine Terms and Conditions